Crops ›› 2017, Vol. 33 ›› Issue (2): 51-58.doi: 10.16035/j.issn.1001-7283.2017.02.009

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Cloning and Sequence Analysis of GmWRI1a Gene Promoter in Soybean

Yan Li,Yang Qiang,Shao Yupeng,Li Dandan,Wang Zhikun,Li Wenbin   

  1. Northeast Agricultural University/Key Laboratory of Soybean Biology, Chinese Ministry of Education/Key Laboratory of Soybean Biology and Genetics Breeding,Chinese Agriculture Ministry,Harbin 150030,Heilongjiang,China
  • Received:2016-11-21 Revised:2017-02-26 Online:2017-04-15 Published:2018-08-26
  • Contact: Zhikun Wang,Wenbin Li

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Abstract:

According to the Williams 82’s genomic sequences reported in the Phytozome, primers were designed by Primer Premier 5.0. The promoter product sequence of GmWRI1a gene was amplified by PCR method. The recombinant vectors pGM-T-pGmWRI1a and PBI121-pGmWRI1a were constructed. The positive clones were identified by PCR amplification and restriction enzyme digestion. The full length of the GmWRI1a promoter was 1 687bp. The promoter sequence contained the necessary initiation transcription sites, such as TATA-box,CTTA-box and multiple other cis-acting elements: light responsiveness element, gibberellin response element, cis-acting regulatory element related to meristem expression, drought-inducing elements, and others. Cloning and characterization of the GmWRI1a gene promoter will provide basis for the further study of regulation and functional analysis of GmWRI1a gene in soybean.

Key words: Promoter of GmWRI1a gene, Soybean, Sequence analysis, Cis-regulatory element

Fig.1

DNA extract result of Dongnong 47"

Fig.2

PCR results of GmWRI1a gene promoter"

Fig.3

PCR identify of pGM-T-pGmWRI1a recombinant plasmid"

Fig.4

Sequence comparison result of GmWRI1a gene promoter"

Fig.5

GmWRI1a gene promoter sequential analysis The possible cis-acting elements in the sequence is signed by frames; The black is transcriptional start site;start codon ATG is signed by the double underline"

Table 1

GmWRI1a gene promoter cis-form regulatory sequence position and function"

调控元件Regulatory sequence 位置Position 核心序列Core sequence 预测功能Function
ACE
 797（+）
 ACGTGGA
 光应答元件
Cis-acting element involved in light responsiveness
AE-Box

 176（+）,
1278（+）,
655（–）		 AGAAAC

 光响应模块
Part of a module for light response
ATC-motif
 1227（–）
 AGTAATCT
 光应答元件
Part of a conserved DNA module involved in light responsiveness
CATT-motif
 210（+）
 GCATTC
 光响应一部分
Part of a light responsive
GA-motif
 835（–）
 AAAGATGA
 光响应元件
Part of a light responsive element
G-box
 795（+）,
796（–）		 TGACGTGG
CACGTC		 光反应调节元件
Cis-acting regulatory element involved in light responsiveness
调控元件Regulatory sequence 位置Position 核心序列Core sequence 预测功能Function
GT1-motif

 674（+）,
1056（+）,
914（–）		 GGTTAA

 光反应元件
Light responsive element
CTA-box
 1008（+）
 GCCACT
 表达分生组织相关
Cis-acting regulatory element related to meristem expression
P-Box
 896（–）
 CCTTTTG
 赤霉素应答元件
Gibberellin-responsive element
GARE-motif
 603（+）,
1482（+）		 AAACAGA
TCTGTTG		 赤霉素响应元件
Gibberellin-responsive element
CE3
 802（+）
 GACGCGTGT
 ABA和VP1响应元件
Cis-acting element involved in ABA and VP1 responsiveness
MBS
 791（+）
 TAACTG
 MYB参与抗旱诱导结合位点
MYB binding site involved in drought-inducibility
Skn-1_motif
 362（+）,
664（+）		 GTCAT
 胚乳表达所需的调节元件
Cis-acting regulatory element required for endosperm expression
GCN4_motif

 370（+）,
1005（+）,
818（+）		 TGAGTCA

 胚乳表达顺式调控元件
Cis-regulatory element involved in endosperm expression

Fig.6

PCR identify of pBI-pGmWRI1a recombinant plasmid"

Fig.7

Hind Ⅲ、XbaⅠenzyme identify of pBI-pGmWRI1a recombinant plasmid"

[1] Pabo C O, Sauer R T . Transcription factors structural families and principles of DNA recognition.Annual Review of Biochemistry. 1992,61:1053-1095.
doi: 10.1146/annurev.bi.61.070192.005201 pmid: 1497306
[2] 姜显光 . 植物油脂中脂肪酸的分析研究. 大连:辽宁师范大学, 2008.
doi: 10.7666/d.y1341481
[3] Durrett T P, Benning C, Ohlrogge J . Plant triacylglycerols as feedstocks for the production of biofuel. Plant Journal, 2008,54:593-607.
doi: 10.1111/j.1365-313X.2008.03442.x pmid: 18476866
[4] Thelen J J, Ohlrogge J B . Metabolic engineering of fatty acidbiosynthesis in plants. Metabolic Engineering, 2002,4(1):12-21.
doi: 10.1006/mben.2001.0204 pmid: 11800570
[5] Abogadallah G M, Nada R M, Malinowski R , et al. Overexpression of HARDY,an AP2/ERF gene from arabidopsis,improves drought and salt tolerance by reducing transpiration and sodium uptake in transgenic Trifolium alexandrinum L. Planta, 2011,233(6):1265-1276.
doi: 10.1007/s00425-011-1382-3
[6] Joo J, Choi H J, Lee Y H , et al. A transcriptional repressor of the ERF family confers drought tolerance to rice and regulates genes preferentially located on chromosome 11. Planta, 2013,238(1):155-170.
doi: 10.1007/s00425-013-1880-6
[7] Wei M, Que K, Vincent A , et al. Wrinkled1a ubiquitous regulator in oil accumulating tissues from arabidopsis embryos to oil palm Mesocarp. Plos One, 2013,8(7):e68887 .
doi: 10.1371/journal.pone.0068887
[8] O’Hara P, SlabasA R, Fawcett T . Fatty acid and lipid biosynthetic genes are expressed at constant molar ratios but different absolute levels during embryogenesis. Plant Physiology, 2002,129:310-320.
doi: 10.1104/pp.010956
[9] NapierJ A , GrahamA. Tailoring plant lipid composition:designer oilseeds come of age. Current Opinion in Plant Biology, 2010,13:330-337.
doi: 10.1016/j.pbi.2010.01.008 pmid: 20185359
[10] 韩志萍, 安利佳, 侯和胜 . AP2/EREBP转录因子的结构与功能.中国农学通报, 2006(3):33-38.
doi: 10.3969/j.issn.1000-6850.2006.03.009
[11] 默韶京 . 长穗偃麦草中AP2/EREBP类转录因子基因的克隆与功能验证. 保定:河北农业大学, 2011.
doi: 10.7666/d.y1897324
[12] Toshitsugu N, Kaoru S, Tatsuhito F , et al. Genome-wide analysis of the ERF gene family in arabidopsis and rice. Plant Physiology, 2006,140(2):411-432.
doi: 10.1104/pp.105.073783 pmid: 16407444
[13] Weigel D . The APETALA2 domain is related to a novel type of DNA binding domain. Plant Cell, 1995,7(4):388-389.
doi: 10.2307/3870077 pmid: 7773013
[14] Moose S P, Sisco P H . Glossy15,an APETALA2-like gene from maize that regulates leaf epidermal cell identity. Genes & Development, 1996,10(23):3018-3027.
doi: 10.1101/gad.10.23.3018 pmid: 8957002
[15] Gu Y Q, Wildermuth M C, Chakravarthy S , et al. Tomato transcription factors Pti4,Pti5 and Pti6 activate defense responses when expressed in arabidopsis. Plant Cell, 2002,14(4):817-831.
doi: 10.1105/tpc.000794
[16] 鲁亚萍, 刘风珍, 万勇善 . 花生转录因子WRI1基因特征的in silico分析.分子植物育种, 2012(3):363-370.
[17] 张经军 . 甘蓝型油菜(Brassica napus)WRI1同源基因克隆及功能初步分析. 石家庄:河北师范大学, 2007.
doi: 10.7666/d.y1099930
[18] 王志坤, 常健敏, 李文滨 . 大豆GmWRI1a基因克隆及生物信息学分析.东北农业大学学报, 2013(7):11-16.
doi: 10.3969/j.issn.1005-9369.2013.07.003
[19] Poxleitner M, Rogers S W, Lacey S A , et al. A role for caleosin in degradation of oil-body storage lipid during seed germination. Plant Journal, 2006,47(6):917-933.
doi: 10.1111/tpj.2006.47.issue-6
[20] 王志坤, 常健敏, 李丹丹 , 等. 大豆GmWRI1a基因克隆及生物信息学分析. 东北农业大学学报, 2013,44(7):11-16.
doi: 10.3969/j.issn.1005-9369.2013.07.003
[21] Ohto M A, Floyd S K, Fischer R L , et al. Effects of APETALA2 on embryo,endosperm and seed coat development determine seed size in arabidopsis. Sexual Plant Reproduction, 2009,22(4):277-289.
doi: 10.1007/s00497-009-0116-1
[22] Kang M, Qian Z, Zhu D , et al. Characterization of micro-RNAs expression during maize seed development. BMC Genomics, 2012,13(1):360.
doi: 10.1186/1471-2164-13-360 pmid: 3468377
[23] 赵利锋, 柴团耀 . AP2/EREBP转录因子在植物发育和胁迫应答中的作用. 植物学报, 2008,25(1):89-101.
doi: 10.3969/j.issn.1674-3466.2008.01.013
[24] Thirugnanasambantham K, Durairaj S, Saravanan S , et al. Role of ethylene response transcription factor (ERF) and its regulation in response to stress encountered by plants. Plant Molecular Biology Reporter, 2015,33(3):1-11.
doi: 10.1007/s11105-014-0732-2
[25] 路静, 赵华燕, 何奕昆 , 等. 高等植物启动子及其应用研究进展. 自然科学进展, 2004,14(8):856-861.
doi: 10.3321/j.issn:1002-008X.2004.08.003
[26] Lau O S, Deng X W . Plant hormone signaling lightens up:integrators of light and hormones. Current Opinion in Plant Biology, 2010,13(5):571-577
doi: 10.1016/j.pbi.2010.07.001 pmid: 20739215
[27] Wu X L, Liu Z H, Hu Z H , et al. BnWRI1 coordinates fatty acid biosynthesis and photosynthesis pathways during oil accumulation in rapeseed. Journal of Integrative Plant Biology, 2014,56(6):582-593.
doi: 10.1111/jipb.v56.6
[28] Surokin L P, Chua N H . Binding sites for two novel phosphoproteins,3AF5 and 3AF3,are required for rhcS-3A expression, Plant Cell, 1992,4(4):473-483.
doi: 10.2307/3869448 pmid: 1498605
[29] 李丹丹, 闫丽, 常健敏 , 等. 大豆GmWRI1基因在糖、植物激素及盐胁迫下的表达分析.作物杂志, 2015(4):41-46.
doi: 10.16035/j.issn.1001-7283.2015.04.010
[30] Jurriaan T, Victor F, Brigitte M M . The multifaceted role of ABA in disease resistance. Trends in Plant Science, 2009,14(6):310-317.
doi: 10.1016/j.tplants.2009.03.006 pmid: 19443266
[31] 陈其军, 安瑞 . 通过同时激活依赖于ABA和不依赖于ABA的逆境胁迫信号转导途径改善拟南芥和烟草的抗逆性. 2004中国植物生理生态学学术研讨会论文摘要汇编, 2004.
[32] Baud S, Mendoza M S, To A , et al. WRINKLED1 specifies the regulatory action of LEAFYCOTYLEDON2 towards fatty acid metabolism during seed maturation in arabidopsis. Plant Journal for Cell & Molecular Biology, 2007,50(5):825-838.
[33] Mu J, Tan H, Zheng Q , et al. LEAFYCOTYLEDON1 is a key regulator of fatty acid biosynthesis in arabidopsis. Plant Physiology, 2008,148(2):1042-1054.
doi: 10.1104/pp.108.126342
[34] Yue J, Sun H, Zhang W , et al. Wheat homologs of yeast ATG6 function in autophagy and are implicated in powdery mildew immunity. BMC Plant Biology, 2015,15(1):95.
doi: 10.1186/s12870-015-0472-y pmid: 25888209
[35] Andre C, Froehlich J, Moll M , et al. A heteromeric plastidic pyruvate kinase complex involved in seed oil biosynthesis in arabidopsis. Plant Cell, 2007,19(6):2006-2022.
doi: 10.1105/tpc.106.048629 pmid: 17557808
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